基于GC-MS代谢组学技术对叶酸调控鸡原代肝细胞代谢的研究

doi:10.11843/j.issn.0366-6964.2018.09.013

摘要/Abstract

摘要：

本试验旨在研究叶酸对鸡原代肝细胞代谢的影响，为探究叶酸调控家禽肝代谢的机制提供参考，也为其在家禽养殖上的合理有效应用提供理论基础和科学依据。选择新生雏鸡分离得到肝细胞，待细胞长至铺满板壁80%左右时更换处理培养基，试验分为对照组和叶酸添加组，每个处理6个重复，处理12 h后，收集细胞测定细胞内三酰甘油、游离脂肪酸（NEFA）和胆固醇含量，并采用GC-MS技术进行代谢组检测，建立OPLS-DA模型，筛选两组的差异代谢物，并基于KEGG数据库对差异代谢物进行代谢通路分析。结果表明，与对照组相比，叶酸的添加显著降低了细胞内的三酰甘油和胆固醇含量（P<0.05），显著提高了游离脂肪酸的水平（P<0.05），除此之外，共鉴定到23个上调和9个下调的代谢物，主要为小分子的糖、酸以及氨基酸等，富集到31条通路上，包括氨基酸类代谢、糖类代谢和脂类代谢等。综上所述，叶酸促进鸡原代肝细胞内的脂类分解，参与调控细胞内的糖、脂以及氨基酸代谢，为今后探究叶酸调控家禽代谢机制提供参考。

Abstract:

The study was conducted to investigate the effect of folic acid on metabolism of chicken primary hepatocytes, aiming to provide a reference for exploring regulation mechanism of folic acid on hepatic metabolism and also provide theoretical and scientific basis for its application in poultry. New-born chickens were used for separating primary liver cells. When the confluence reached to about 80%, hepatocytes were exposed to treatment medium. The control group and folic acid addition group were contained in the study. There were 6 replicates for each group. After 12 h treatment, cells were collected to detect triglyceride, NEFA and cholesterols contents. Metabolomics analysis was carried out by GC-MS method. The OPLS-DA model was established to identify differential metabolites, and pathways enriched based on differential metabolites were analyzed through KEGG system. The results showed that folic acid significantly decreased triglyceride and cholesterol contents in hepatocytes compared with the control group (P<0.05). NEFA level was higher in folic acid addition group (P<0.05). Twenty three up-regulation and 9 down-regulation metabolites were found between two groups, which mainly included amino acid, small molecule sugar and acid. Thirty one pathways were enriched based on differential metabolites containing amino acid metabolism, carbohydrate metabolism, lipid metabolism, and so on. The results indicated that folic acid could promote lipid hydrolysis in chicken primary hepatocytes, and was involved in regulating sugar, lipid and amino acid metabolism, which would provide a reference for exploring regulatory mechanism of folic acid on hepatic metabolism in future.

中图分类号:

S831.5

刘艳利, 孙文强, 韩迪, 段玉兰, 杨欣, 杨小军. 基于GC-MS代谢组学技术对叶酸调控鸡原代肝细胞代谢的研究[J]. 畜牧兽医学报, 2018, 49(9): 1919-1927.

LIU Yan-li, SUN Wen-qiang, HAN Di, DUAN Yu-lan, YANG Xin, YANG Xiao-jun. The Study on Folic Acid Regulating Metabolism of Chicken Primary Hepatocytes by GC-MS Metabolomics[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(9): 1919-1927.

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